Association of tyrosine hydroxylase 01 (TH01) microsatellite and insulin gene (INS) variable number of tandem repeat (VNTR) with type 2 diabetes and fasting insulin secretion in Mexican population.

PURPOSE: A variable number of tandem repeats (VNTR) in the insulin gene (INS) control region may be involved in type 2 diabetes (T2D). The TH01 microsatellite is near INS and may regulate it. We investigated whether the TH01 microsatellite and INS VNTR, assessed via the surrogate marker single nucleotide polymorphism rs689, are associated with T2D and serum insulin levels in a Mexican population. METHODS: We analyzed a main case-control study (n = 1986) that used univariate and multivariate logistic regression models to calculate the risk conferred by TH01 and rs689 loci for T2D development; rs689 results were replicated in other case-control (n = 1188) and cross-sectional (n = 1914) studies. RESULTS: TH01 alleles 6, 8, 9, and 9.3 and allele A of rs689 were independently associated with T2D, with differences between sex and age at diagnosis. TH01 alleles with ≥ 8 repeats conferred an increased risk for T2D in males compared with ≤ 7 repeats (odds ratio, ≥ 1.46; 95% confidence interval, 1.1-1.95). In females, larger alleles conferred a 1.5-fold higher risk for T2D when diagnosed ≥ 46 years but conferred protection when diagnosed ≤ 45 years. Similarly, rs689 allele A was associated with T2D in these groups. In males, larger TH01 alleles and the rs689 A allele were associated with a significant decrease in median fasting plasma insulin concentration with age in T2D cases; the reverse occurred in controls. CONCLUSION: Larger TH01 alleles and rs689 A allele may potentiate insulin synthesis in males without T2D, a process disabled in those with T2D.

In vitro screening of technical lignins to determine their potential as hay preservatives.

Our objectives were to evaluate technical lignins for their antifungal properties against 3 molds and 1 yeast causing hay spoilage, and their ability to preserve ground high-moisture alfalfa hay nutritive value in vitro. In experiment 1, 8 technical lignins and propionic acid (PRP; positive control) were tested at a dose of 40 mg/mL. The experiment had a randomized complete block design (RCBD, 4 runs) and a factorial arrangement of 3 molds × 10 additives (ADV). The effects of the ADV on yeast were evaluated separately with a RCBD. Sodium lignosulfonate (NaL) and PRP were the only treatments with 100 ± 2.8% inhibition of fungi. In experiment 2, the minimum inhibitory concentration (MIC) for selected lignins and PRP were determined. At pH 4, NaL had the lowest MIC across the molds (20-33.3 mg/mL) and magnesium lignosulfonate (MgL) for the yeast (26.7) among the lignins. However, PRP had MIC values that were several-fold lower across all fungi (1.25-3.33). In experiment 3, a RCBD (5 blocks) with a 3 (ADV; NaL, MgL, and PRP) × 4 (doses: 0, 0.5, 1, and 3% wt/wt fresh basis) factorial arrangement of treatments was used to evaluate the preservative effects of ADV in ground high-moisture alfalfa hay inoculated with a mixture of the fungi previously tested and incubated under aerobic conditions in vitro. After 15 d, relative to untreated hay (14.9), dry matter (DM) losses were lessened by doses as low as 1% for NaL (3.39) and 0.5% for PRP (0.81 ± 0.77%). The mold count was reduced in both NaL at 3% (3.92) and PRP as low as 0.5% (3.94) relative to untreated hay (7.76 ± 0.55 log cfu/fresh g). Consequently, sugars were best preserved by NaL at 3% (10.1) and PRP as low as 0.5% (10.5) versus untreated (7.99 ± 0.283% DM), while keeping neutral detergent fiber values lower in NaL (45.9) and PRP-treated (45.1) hays at the same doses, respectively, relative to untreated (49.7 ± 0.66% DM). Hay DM digestibility was increased by doses as low as 3% for NaL (67.5), 1% MgL (67.0), and 0.5% PRP (68.5) versus untreated hay (61.8 ± 0.77%). The lowest doses increasing neutral detergent fiber digestibility relative to untreated hay (23.3) were 0.5% for MgL and PRP (30.5 and 30.1, respectively) and 1% for NaL (30.7 ± 1.09% DM). Across technical lignins, NaL showed the most promise as a potential hay preservative. However, its effects were limited compared with PRP at equivalent doses. Despite not having an effect on preservation, MgL improved DM digestibility by stimulating neutral detergent fiber digestibility. This study warrants further development of NaL under field conditions.

Symposium review: Technologies for improving fiber utilization.

The forage lignocellulosic complex is one of the greatest limitations to utilization of the nutrients and energy in fiber. Consequently, several technologies have been developed to increase forage fiber utilization by dairy cows. Physical or mechanical processing techniques reduce forage particle size and gut fill and thereby increase intake. Such techniques increase the surface area for microbial colonization and may increase fiber utilization. Genetic technologies such as brown midrib mutants (BMR) with less lignin have been among the most repeatable and practical strategies to increase fiber utilization. Newer BMR corn hybrids are better yielding than the early hybrids and recent brachytic dwarf BMR sorghum hybrids avoid lodging problems of early hybrids. Several alkalis have been effective at increasing fiber digestibility. Among these, ammoniation has the added benefit of increasing the nitrogen concentration of the forage. However, few of these have been widely adopted due to the cost and the caustic nature of the chemicals. Urea treatment is more benign but requires sufficient urease and moisture for efficacy. Ammonia-fiber expansion technology uses high temperature, moisture, and pressure to degrade lignocellulose to a greater extent than ammoniation alone, but it occurs in reactors and is therefore not currently usable on farms. Biological technologies for increasing fiber utilization such as application of exogenous fibrolytic enzymes, live yeasts, and yeast culture have had equivocal effects on forage fiber digestion in individual studies, but recent meta-analyses indicate that their overall effects are positive. Nonhydrolytic expansin-like proteins act in synergy with fibrolytic enzymes to increase fiber digestion beyond that achieved by the enzyme alone due to their ability to expand cellulose microfibrils allowing greater enzyme penetration of the cell wall matrix. White-rot fungi are perhaps the biological agents with the greatest potential for lignocellulose deconstruction, but they require aerobic conditions and several strains degrade easily digestible carbohydrates. Less ruminant nutrition research has been conducted on brown rot fungi that deconstruct lignocellulose by generating highly destructive hydroxyl radicals via the Fenton reaction. More research is needed to increase the repeatability, efficacy, cost effectiveness, and on-farm applicability of technologies for increasing fiber utilization.

The current prevalence of asthma, allergic rhinitis, and eczema related symptoms in school-aged children in Costa Rica.

OBJECTIVE: Asthma prevalence in Costa Rica is among the highest worldwide. We aimed to determine the prevalence of asthma among school-age children in the Central Highland Area of Costa Rica. METHODS: Cross-sectional study using the International Study of Asthma and Allergies in Childhood (ISAAC) questionnaire was performed. Parents or guardians of children aged 6-13 years completed written questionnaires. RESULTS: Total of 2817 school-aged children returned these questionnaires (74.1% return rate). The prevalence of asthma, rhinitis, and eczema was 21.9%, 42.6%, and 19.2%, respectively. The co-existence of the 3 diseases was seen in 22.6% of children with asthma. Boys had a slightly higher prevalence of these conditions, and younger children had higher prevalence of asthma and eczema, but lower prevalence of rhinitis than older children. The use of acetaminophen and antibiotics in the first 12 months of life showed a significant association with the prevalence of asthma, rhinitis, and eczema. Wheezing with exercise, dry cough at night, and ever rhinitis was highly associated with asthma symptoms in the last 12 months. In contrast, no association was found between children exposed to smoking at home. Frequent traffic next to the house was reported more frequently by the parents of children with asthma, although no significant association was found. CONCLUSION: The prevalence of asthma showed a significant decrease compared to previous studies. However, there was an unexpected high prevalence of rhinitis. Exposure to acetaminophen and antibiotic during the first year of life was highly associated with asthma symptoms.

Effects of homolactic bacterial inoculant on the performance of lactating dairy cows.

The objective of this experiment was to determine the effect of applying a homofermentative bacterial inoculant to corn silage on the performance of dairy cows. After harvesting, corn forage was treated with nothing (CON) or with an inoculant containing a mixture of Lactococcus lactis, Lactobacillus plantarum, and Enterococcus faecium at 1.5 × 105 cfu/g of fresh forage (MC; SiloSolve MC, Chr. Hansen A/S, Hørsholm, Denmark). After 186 d of storage in Ag-Bags (A Miller-St. Nazianz Inc., St. Nazianz, WI), silages were fed as part of a total mixed ration containing 55% concentrates, 10% alfalfa hay, and 35% CON or MC corn silage. Sixty early-lactation Holstein dairy cows (30 multiparous and 30 primiparous) housed in a freestall barn with Calan gates (American Calan Inc., Northwood, NH) were assigned to the dietary treatments from 20 to 100 d in milk. Silage inoculated with MC had a more homofermentative pattern evidenced by greater lactic acid concentration (3.83 vs. 4.48% of DM) and lower concentrations of acetic (2.34 vs. 1.68% of DM) and propionic (0.37 vs. 0.10% of DM) acids and ammonia (9.11 vs. 7.82% of N) for CON and MC, respectively. Dry matter intake (23.1 vs. 23.2 kg/d) did not differ among treatments, but the MC silage had greater apparent digestibility of DM (68.8 vs. 70.8%), which led to greater yields of milk (37.7 vs. 38.5 kg/d), fat-corrected milk (37.6 vs. 38.4 kg/d), milk fat (1.30 vs. 1.33 kg/d), and lactose (1.83 vs. 1.92 kg/d) for CON and MC cows, respectively. Milk from cows fed MC silage had higher lactose (4.86 vs. 4.93%), lower protein (2.93 vs. 2.83%), and similar contents of fat (3.47 vs. 3.44%) compared with CON cows. Feed efficiency (fat-corrected milk/dry matter intake) was not affected by treatment (1.69 vs. 1.72 for CON and MC, respectively). Inoculation of corn silage with the homofermentative inoculant increased digestibility of the total mixed ration and increased milk yield by lactating dairy cows.

Bacterial and fungal communities, fermentation, and aerobic stability of conventional hybrids and brown midrib hybrids ensiled at low moisture with or without a homo- and heterofermentative inoculant.

We evaluated the effects of adding a combination inoculant to 4 corn (Zea mays L.) hybrids harvested at low moisture on the nutritive value, fermentation profile, aerobic stability, bacterial and fungal populations, and community structure. The treatment design was the factorial combination of 4 corn hybrids ensiled with (INO) and without (CON) inoculant. The hybrids were TMF2R737 (MCN), F2F817 (MBR), P2089YHR (PCN), and PI144XR (PBR), ensiled at 44.0, 38.1, 42.0, and 41.3% of dry matter, respectively; MBR and PBR were brown midrib mutants. The inoculant contained Lactobacillus buchneri and Pediococcus pentosaceus (4 × 105 and 1 × 105 cfu/g of fresh corn). The experimental design was a complete randomized design with treatments replicated 6 times. Corn was chopped, treated or not with inoculant, packed into 7.6-L bucket silos, and stored for 100 d. At d 0, we found higher bacterial observed operational taxonomic units in the brown midrib mutants (MBR and PBR) relative to MCN and PCN (654 and 534 vs. 434 and 444 ± 15.5, respectively). The bacterial and fungal families with the highest relative abundance (RA) were Enterobacteriaceae (61.4%) and incertae sedis Tremellales (12.5%). At silo opening, we observed no effects of INO treatment on dry matter recovery (∼94.3 ± 1.07%), but aerobic stability was extended for all INO-treated hybrids (∼217 vs. ∼34.7 h), except for MBR (∼49 ± 38 h), due to a decreased yeast population (3.78 vs. 5.13 ± 0.440 log cfu/g of fresh corn) and increased acetic acid concentration (1.69 vs. 0.51 ± 0.132%) compared with the control. Furthermore, INO treatment reduced bacterial (61.2 vs. 276 ± 8.70) and increased fungal (59.8 vs. 43.6 ± 2.95) observed operational taxonomic units compared with CON. We observed that INO treatment increased the RA of Lactobacillaceae across all hybrids (∼99.1 vs. ∼58.9), and to larger extent MBR (98.3 vs. 34.3 ± 5.29), and decreased Enterobacteriaceae (0.614 vs. 23.5 ± 2.825%) among 4 other bacterial families relative to CON. For fungi, INO treatment increased the RA of Debaryomycetaceae (63.1 vs. 17.3 ± 8.55) and 5 other fungal families and decreased the RA of Pichiaceae (6.47 vs. 47.3 ± 10.95) and incertae sedis Saccharomycetales (8.47 vs. 25.9 ± 5.748) compared with CON. The bacterial and fungal community structures changed, due to ensiling, to a distinct and more stable community dominated by Lactobacillaceae and Debaryomycetaceae, respectively, when INO treatment was applied relative to CON. In conclusion, the INO treatment used in this study improved low-moisture whole-crop corn silage quality because of a shift in the bacterial and fungal community composition during ensiling.

The capacity of silage inoculant bacteria to bind aflatoxin B1 in vitro and in artificially contaminated corn silage.

The objectives were to examine the aflatoxin B1 (AFB1)-binding capacity of silage bacteria and factors affecting the responses. Experiments 1 and 2 examined the effects of bacterial strain and population on the AFB1-binding capacity of 10 bacteria. When applied at 106 cfu/mL to an in vitro medium, only Lactobacillus plantarum PT5B bound the AFB1 and the binding capacity was low (4%). When applied at 109 cfu/mL, all 10 bacteria bound AFB1, but L. plantarum R2014 (Lp) and EQ12, Lactobacillus buchneri R1102 (Lb), and Pediococcus acidilactici R2142 and EQ01 (Pa) had the greatest capacity (23.9 to 33%). Experiment 3 examined the AFB1-binding capacity of viable and nonviable (HCl-treated) forms of Lp, Lb, and Pa at different pH. Nonviable Lb and Lp, but not Pa, increased AFB1 binding. Binding of AFB1 was greatest at pH 2.5 and least at pH 8. As the nonviable Lb and Lp that bound AFB1 in experiment 3 would not be effective silage inoculants, experiment 4 examined effects of benign versus severe treatments (85 vs. 100°C; pH 2.5 vs. <1) on the viability of Lp, Lb, and Pa. The population of bacteria was reduced from 9 to 4 log cfu/mL by treatment with HCl at pH 2.5 and to 2 log cfu/mL by 85 or 100°C, whereas acidification at pH <1 eliminated the bacteria. Experiment 5 determined the effect of the ensiling duration and benign treatment methods [37 (viable cells) or 85°C (heated cells) or acidification with HCl at pH 2.5 (acid-treated cells)] on binding of AFB1 and silage quality during the fermentation of corn forage. Corn forage was ensiled after treatment with only deionized water (control), AFB1 (30 µg/kg of fresh forage), or a mixture of AFB1 and 109 cfu/g of each of the treated bacteria. Adding AFB1 alone to corn forage reduced the pH decline during the first 3 d of ensiling and increased or tended to increase butyric acid concentration and final pH after ensiling for 21 d. Bacterial inoculation inhibited these negative effects. The fermentation profile of silage treated with Lb and Pa did not differ from those of the control silage. In all silages treated with the toxin, the AFB1 concentration decreased linearly (from 30 to ≤0.35 µg/kg) within 3 d of ensiling. Certain silage bacteria can bind AFB1 but the efficacy depends on several factors.

Laboratory silo type and inoculation effects on nutritional composition, fermentation, and bacterial and fungal communities of oat silage.

The objectives were to evaluate (1) the use of 2 types of experimental silos (S) to characterize whole-crop oat (Avena sativa L.) silage with or without addition of an inoculant (I), and (2) the effect of inoculation on the microbial community structure of oats ensiled using only plastic bucket silos (BKT). From each of 6 sections in a field, oats were harvested, treated (INO) or not (CON) with inoculant, packed into 19-L BKT or vacuum bags (BG), and ensiled for 217 d. The inoculant added contained Lactobacillus buchneri and Pediococcus pentosaceus (4 × 105 and 1 × 105 cfu/g of fresh oats, respectively). The experimental design was a complete randomized design replicated 6 times. Treatment design was the factorial combination of 2 S × 2 I. Some differences existed between BG versus BKT at silo opening (217 d), including a decreased CP (7.73 vs. 7.04 ± 0.247% of DM) and ethanol (1.93 vs. 1.55 ± 0.155) and increased lactic acid (4.28 vs. 3.65 ± 0.241), respectively. Also, WSC and mold counts were reduced in BG versus BKT for CON (1.78 vs. 2.70 ± 0.162% of DM and 0.8 vs. 2.82 ± 0.409 log cfu/fresh g) but not for INO (∼1.53 and 1.55), respectively. Application of INO increased DM recovery (96.1 vs. 92.9 ± 0.63%), aerobic stability (565 vs. 133 ± 29.2 h), acetic acid (2.38 vs. 1.22 ± 0.116% of DM), and reduced NDF (65.0 vs. 67.0 ± 0.57), ADF (36.7 vs. 38.1 ± 0.60), ethanol (0.63 vs. 2.85 ± 0.155), and yeast counts (1.10 vs. 4.13 ± 0.484 log cfu/fresh g) in INO versus CON, respectively. At d 0, no differences were found for S and I on the nutritional composition and background microbial counts. Leuconostocaceae (82.9 ± 4.27%) and Enterobacteriaceae (15.2 ± 3.52) were the predominant bacterial families and unidentified sequences were predominant for fungi. A higher relative abundance of the Davidiellaceae fungal family (34.3 vs. 19.6 ± 4.47) was observed in INO versus CON. At opening (217 d), INO had a lower relative abundance of Leuconostocaceae (42.3 vs. 95.8 ± 4.64) and higher Lactobacillaceae (57.4 vs. 3.9 ± 4.65) versus CON. Despite several differences were found between BKT and BG, both techniques can be comparable for characterizing effects of INO on the most basic measures used in silage evaluation. The use of inoculant improved oat silage quality partially by a shift in the bacterial community composition during ensiling, which mainly consisted of an increased relative abundance of Lactobacillaceae and reduction of Leuconostocaceae relative to CON.

Effects of maturity at ensiling of bermudagrass and fibrolytic enzyme application on the performance of early-lactation dairy cows.

The objective of this study was to examine effects of adding fibrolytic enzymes to diets containing bermudagrass ensiled after 4 or 7wk of regrowth on the diet digestibility, ruminal fermentation and performance of lactating cows, and the interaction of the treatments. In experiment 1, 64 Holstein cows (22±4d in milk) were assigned to an experiment with a 2×2 factorial treatment arrangement and a 56-d duration. Treatments were diets containing 4 or 7wk regrowth bermudagrass silage without or with an exogenous fibrolytic enzyme cocktail. The cellulase-xylanase enzyme was applied at 2.33g/kg of total mixed ration dry matter (DM) during mixing immediately before feeding. Experiment 2 was aimed at examining treatment effects on the ruminal fermentation profile. Four ruminally cannulated cows were assigned to the 4 treatments using a 4×4 Latin square design with 14-d periods. No enzyme by maturity interaction was detected for any measurement. Regardless of forage maturity, applying the fibrolytic enzyme did not affect DM intake, milk yield, apparent digestibility, feed efficiency, energy balance, and ruminal fermentation though it tended to increase milk lactose concentration (4.88 vs. 4.81%). Feeding the 4-wk diet instead of the 7-wk diet increased DM intake (22.4 vs. 21.3kg/d), digestibility of DM, neutral detergent fiber, and acid detergent fiber, and tended to increase 3.5%-fat corrected milk yield (47.2 vs. 44.3kg/d) and milk fat yield (1.88 vs. 1.73kg/d). Therefore, daily intake of net energy and secretion of milk energy were greater for the 4-wk diet. In addition, the 4-wk diet increased the ruminal concentrations of acetate, propionate, valerate, lactate, and total volatile fatty acids, and decreased ruminal pH, without affecting the acetate:propionate ratio. Feeding fibrolytic enzymes did not improve the performance of early-lactation dairy cows, but harvesting the forage earlier tended to improve milk production.

Improving the performance of dairy cattle with a xylanase-rich exogenous enzyme preparation.

The objective of this experiment was to examine effects of adding 2 exogenous fibrolytic enzymes (EFE) to the total mixed ration (TMR) on the performance of lactating dairy cows (experiment 1) and the kinetics of ruminal degradation of the diet (experiment 2). Twelve EFE had been screened in a series of in vitro assays that identified the most potent EFE and their optimal doses for increasing the digestibility of bermudagrass. In experiment 1, 66 Holstein cows (21±5 d in milk) were grouped by previous milk production and parity (45 multiparous and 21 primiparous) and assigned randomly to 1 of the following 3 treatments: (1) control (CON, untreated), (2) Xylanase Plus [2A, 1mL/kg of TMR dry matter (DM); Dyadic International, Jupiter, FL], and (3) a 75:25 (vol/vol) mixture of Cellulase Plus and Xylanase Plus EFE (3A, 3.4mL/kg of TMR DM; Dyadic International). The EFE were sprayed twice daily onto a TMR (10% bermudagrass silage, 35% corn silage, 5% alfalfa-orchardgrass hay mixture, and 50% concentrates; DM basis) and fed for a 14-d training and covariate period and a 70-d measurement period. Experiment 2 aimed to examine the in situ DM ruminal degradability and ruminal fermentation measurements of the diets fed in experiment 1. Three ruminally fistulated lactating Holstein cows were assigned to the diets. The experiment had a 3×3 Latin square design with 23-d periods. In experiment 1, application of 2A increased intakes (kg/d) of DM (23.5 vs. 22.6), organic matter (21.9 vs. 20.9), and crude protein (3.9 vs. 3.7) and tended to increase yields (kg/d) of fat-corrected milk (41.8 vs. 40.7) and milk fat (1.48 vs. 1.44). In particular, 2A increased milk yield (kg/d) during wk 3 (41.2 vs. 39.8, tendency), 6 (41.9 vs. 40.1), and 7 (42.1 vs. 40.4), whereas 3A increased milk yield (kg/d) during wk 6 (41.5 vs. 40.1, tendency), 8 (41.8 vs. 40.0), and 9 (40.9 vs. 39.5, tendency). In experiment 2, EFE treatment did not affect ruminal DM degradation kinetics or ruminal pH, ammonia-N, and volatile fatty acid concentration. Application of 2A to the bermudagrass-based TMR increased DM intake and milk production, implying that this EFE could be used to increase the performance of lactating dairy cows fed diets containing up to 10% bermudagrass.

Panniculitis caused by Mycobacterium monacense mimicking erythema induratum: a case in Ecuador.

We report the first case of recently characterized species M. monacense associated with chronic nodular vasculitis, infecting a young woman. This case represents the first isolation of M. monacense from Ecuador. The isolate was identified by conventional and molecular techniques.

Low thermal conductivity and improved thermoelectric performance of nanocrystalline silicon germanium films by sputtering.

Si x Ge1-x alloys are well-known thermoelectric materials with a high figure of merit at high temperatures. In this work, metal-induced crystallization (MIC) has been used to grow Si0.8Ge0.2 films that present improved thermoelectric performance (zT = 5.6 × 10(-4) at room temperature)--according to previously reported values on films--with a relatively large power factor (σ · S (2) = 16 µW · m(-1) · K(-2)). More importantly, a reduction in the thermal conductivity at room temperature (κ = 1.13 ± 0.12 W · m(-1) · K(-1)) compared to other Si-Ge films (∼3 W · m(-1) · K(-1)) has been found. Whereas the usual crystallization of amorphous SiGe (a-SiGe) is achieved at high temperatures and for long times, which triggers dopant loss, MIC reduces the crystallization temperature and the heating time. The associated dopant loss is thus avoided, resulting in a nanostructuration of the film. Using this method, we obtained Si0.8Ge0.2 films (grown by DC plasma sputtering) with appropriate compositional and structural properties. Different thermal treatments were tested in situ (by heating the sample inside the deposition chamber) and ex situ (annealed in an external furnace with controlled conditions). From the studies of the films by: x-ray diffraction (XRD), synchrotron radiation grazing incidence x-ray diffraction (SR-GIXRD), micro Raman, scanning electron microscopy (SEM), x-ray photoemission spectroscopy (XPS), Hall effect, Seebeck coefficient, electrical and thermal conductivity measurements, we observed that the in situ films at 500 °C presented the best zT values with no gold contamination.

Effect of adding cofactors to exogenous fibrolytic enzymes on preingestive hydrolysis, in vitro digestibility, and fermentation of bermudagrass haylage.

Our objectives were to examine if adding metal ion cofactors (COF) to exogenous fibrolytic enzymes (EFE) would increase the beneficial effects of the EFE on the preingestive hydrolysis and in vitro digestibility and fermentation of bermudagrass haylage. In experiment 1, 5 COF (Mn(2+), Co(2+), Fe(2+), Ca(2+), and Mg(2+)) were screened to select the best candidates for synergistically enhancing release of water-soluble carbohydrates (WSC) from bermudagrass haylage by 5 EFE. The 5 EFE (1A, 2A, 11C, 13D, and 15D) were sourced from Trichoderma reesei and Aspergillus oryzae and they were the most effective of 12 EFE at increasing the neutral detergent fiber digestibility of bermudagrass haylage in a previous trial. Adding 1mM of each of the COF to EFE 2A or 11C synergistically increased release of WSC from bermudagrass haylage, as did adding (1mM) Fe(2+) to 1A, Mn(2+), Co(2+), or Fe(2+) to 13D, or Co(2+)or Fe(2+) to 15D. The greatest release of WSC responses were obtained by adding Mn(2+) to 11C (38%) or by adding Fe(2+) to 2A or 13D (10 and 21.9%, respectively). In experiment 2, the effect of increasing the COF dose on in vitro digestibility and fermentation of bermudagrass haylage was examined using the best EFE-COF combinations from experiment 1. Effects of adding increasing doses of these COF on EFE-mediated changes in vitro digestibility depended on the COF-EFE combination. Adding 10mM Mn(2+) alone to bermudagrass haylage increased DMD and NDFD by 2.7 and 6.3% and adding 11C alone increased these measures by 6.6 and 15.5%, respectively. However, adding 10mM Mn(2+) with 11C resulted in 3.5 and 8.1% increases in DMD and NDFD, respectively, beyond the increases caused by adding 11C alone. Adding Fe(2+) to 2A had no effects on EFE-mediated digestibility responses, but 2A prevented adverse effects of adding Fe(2+) alone on DMD and NDFD. In contrast, adding Fe(2+) to 13D reduced the increases in DMD and NDFD caused by adding the EFE alone. This study shows that adding COF to EFE can synergistically increase, decrease, or not affect the hydrolytic effects of EFE on bermudagrass haylage cell walls. The outcome depends on the specific EFE-COF combination and the COF dose. More research is required to understand the mechanisms resulting in these outcomes to exploit beneficial effects of COF on EFE.

Is the Production of Embryos in Small-Scale Farming an Economically Feasible Enterprise?

The present assay attempts to evaluate the feasibility of using embryo transfer in small community farmers by in vivo study and by modelling the results obtained. From the total of 59 donor cows, 62.7% responded to treatment, with a significant difference (p = 0.002) in the percentage of the response between breeds, being 90.5% (19/21) in Holstein and 47.4% (18/38) in Brahman. A total of 283 embryos were graded as transferable, while 141 as non-transferable, without difference in the percentage of transferable embryo by breed (p = 0.18). The mean of transferable embryos graded as class I and II was not different between Holstein and Brahman (p = 0.96 and p = 0.92, respectively); besides, no differences were observed in the other grades (non-transferable). The highest difference in costs, regardless of its quality by breed, was seen in the lower levels of probable fertility of the embryo transferred, even reaching several hundred dollars. When modelling the expected costs for embryo produced and transferred, values can reach nearly $2000.00 when the probable fertility is only 10%. However, when the probable fertility was 60%, embryo cost was close to $300.00. This technology seems to be viable on average or high-scale systems, having a superovulatory response between 60 and 80% with 4-6 transferrable embryos. Yet, in small-scale farming, due to the reduced number of donors and/or recipients, the costs surpass the economical feasibility of the technique.

Screening exogenous fibrolytic enzyme preparations for improved in vitro digestibility of bermudagrass haylage.

Our objectives were to evaluate the effects of 12 exogenous fibrolytic enzyme products (EFE) on ruminal in vitro neutral detergent fiber digestibility (NDFD) and preingestive hydrolysis of a 4-wk regrowth of bermudagrass haylage (BH), to examine the accuracy of predicting NDFD with EFE activity measures, and to examine the protein composition of the most and least effective EFE at increasing NDFD. In experiment 1, effects of 12 EFE on NDFD of BH were tested. Enzymes were applied in quadruplicate to culture tubes containing ground BH. The suspension was incubated for 24 h at 25 °C before addition of rumen fluid media and further incubation for 24 h at 39 °C. The experiment was repeated twice. In addition, regression relationships between EFE activity measures and NDFD were examined. Compared with the values for the control, 9 EFE-treated substrates had greater NDFD (37.8 to 40.4 vs. 35.6%), 6 had greater total VFA concentration (59.1 to 61.2 vs. 55.4 mM), and 4 had lower acetate-to-propionate ratios (3.03 to 3.16 vs. 3.24). In experiment 2, EFE effects on preingestive fiber hydrolysis were evaluated by incubating enzyme-treated and untreated bermudagrass suspensions in quadruplicate for 24 h at 25 °C and examining fiber hydrolysis measures. Compared with values for the control, 3 EFE reduced neutral detergent fiber concentration (62.8 to 63.7 vs. 67.3%), 10 increased release of water-soluble carbohydrates (26.8 to 58.5 vs. 22.8 mg/g), and 8 increased release of ferulic acid (210 to 391 vs. 198 µg/g). Regression analyses revealed that enzyme activities accurately [coefficient of determination (R(2)) = 0.98] predicted preingestive hydrolysis measures (water-soluble carbohydrates, ferulic acid), moderately (R(2) = 0.47) predicted neutral detergent fiber hydrolysis, but poorly (R(2) ≤ 0.1) predicted dry matter and NDFD. In experiment 3, proteomic tools were used to examine the protein composition of the most and least effective EFE at improving NDFD. Relative to the least effective, the most effective EFE at increasing NDFD contained 10 times more endoglucanase III, 17 times more acetylxylan esterase with a cellulose-binding domain 1, 33 times more xylanase III, 25 times more ß-xylosidase, and 7.7 times more polysaccharide monooxygenase with cellulose-binding domain 1 and 3 times more swollenin. The most effective EFE had a much greater quantity of fibrolytic enzymes and key proteins necessary for hemicellulose and lignocellulase deconstruction. This study identified several EFE that increased the NDFD and in vitro fermentation of 4-wk BH and revealed why some EFE are more effective than others.

Effect of microbial inoculants on the quality and aerobic stability of bermudagrass round-bale haylage.

The objective of this study was to compare the efficacy of using 4 commercially available microbial inoculants to improve the fermentation and aerobic stability of bermudagrass haylage. We hypothesized that the microbial inoculants would increase the fermentation and aerobic stability of the haylages. Bermudagrass (4-wk regrowth) was harvested and treated with (1) deionized water (control); (2) Buchneri 500 (B500; Lallemand Animal Nutrition, Milwaukee, WI) containing 1×10(5) of Pediococcus pentosaceus and 4×10(5) of Lactobacillus buchneri 40788; (3) Biotal Plus II (BPII; Lallemand Animal Nutrition) containing 1.2×10(5) of P. pentosaceus and Propionibacteria freudenreichii; (4) Silage Inoculant II (SI; AgriKing Inc., Fulton, IL) containing 1×10(5) of Lactobacillus plantarum and P. pentosaceus; and (5) Silo King (SK; AgriKing Inc.), containing 1×10(5) of L. plantarum, Enterococcus faecium, and P. pentosaceus, respectively. Forty round bales (8 per treatment; 441±26kg; 1.2×1.2 m diameter) were made and each was wrapped with 7 layers of plastic. Twenty bales were stored for 112 d and the remaining 20 were stored for 30 d and sampled by coring after intermediary storage periods of 0, 3, 7, and 30 d. The pH of control and inoculated haylages sampled on d 3 did not differ. However, B500 and BPII had lower pH (5.77±0.04 vs. 6.16±0.04; 5.06±0.13 vs. 5.52±0.13) than other treatments by d 7 and 30, respectively. At final bale opening on d 112, all treatments had lower pH than the control haylage (4.77±0.07 vs. 5.37±0.07). The B500, BPII, and SI haylages had greater lactic acid and lactic-to-acetic acid ratios than SK and control haylages. No differences were detected in neutral detergent fiber digestibility, dry matter losses, dry matter, lactic and acetic acid concentrations, and yeast and coliform counts. The SK haylage had lower clostridia counts compared with the control (1.19±0.23 vs. 1.99±0.23 cfu/g). Treatments B500, BPII, SI, and SK tended to reduce mold counts and they improved aerobic stability by 236, 197, 188, and 95%, respectively, compared with the control (276±22 vs. 99±22h).

Effect of the dose of exogenous fibrolytic enzyme preparations on preingestive fiber hydrolysis, ruminal fermentation, and in vitro digestibility of bermudagrass haylage.

Our objectives were to evaluate the effects of the dose rates of 5 Trichoderma reesei and Aspergillus oryzae exogenous fibrolytic enzymes (EFE; 1A, 2A, 11C, 13D, and 15D) on in vitro digestibility, fermentation characteristics, and preingestive hydrolysis of bermudagrass haylage and to identify the optimal dose of each EFE for subsequent in vitro and in vivo studies. In experiment 1, EFE were diluted in citrate-phosphate buffer (pH 6) and applied in quadruplicate in each of 2 runs at 0× (control), 0.5×, 1×, 2×, and 3×; where 1× was the respective manufacturer-recommended dose (2.25, 2.25, 10, 15, and 15g of EFE/kg of dry matter). The suspension was incubated for 24h at 25°C and for a further 24h at 39°C after the addition of ruminal fluid. In experiment 2, a similar approach to that in experiment 1 was used to evaluate simulated preingestive effects, except that sodium azide (0.02% wt/vol) was added to the EFE solution. The suspension was incubated for 24h at 25°C and then 15mL of water was added before filtration to extract water-soluble compounds. For both experiments, data for each enzyme were analyzed separately as a completely randomized block design with a model that included effects of EFE dose, run, and their interaction. In experiment 1, increasing the EFE dose rate nonlinearly increased the DM digestibility of 1A, 2A, 11C, and 13D and the neutral detergent fiber digestibility (NDFD) of 1A, 2A, 11C, and 13D. Optimal doses of 1A, 2A, 11C, 13D, and 15D, as indicated by the greatest increases in NDFD at the lowest dose tested, were 2×, 2×, 1×, 0.5×, and 0.5×, respectively. Increasing the dose rate of 2A, 11C, and 13D nonlinearly increased concentrations of total volatile fatty acids and propionate (mM), decreased their acetate-to-propionate ratios and linearly decreased those of samples treated with 1A and 15D. In experiment 2, increasing the dose rate of each EFE nonlinearly decreased concentrations of netural detergent fiber; also, increasing the dose rate of 1A, 2A, 11C, and 1D nonlinearly increased concentrations of water-soluble carbohydrates and free ferulic acid (µg/g). Application of increasing doses of the EFE increased NDF hydrolysis, NDFD, and ruminal fluid fermentation of bermudagrass haylage, but the optimal dose varied with the EFE.

Ruminant Nutrition Symposium: Improving cell wall digestion and animal performance with fibrolytic enzymes.

This paper aimed to summarize published responses to treatment of cattle diets with exogenous fibrolytic enzymes (EFE), to discuss reasons for variable EFE efficacy in animal trials, to recommend strategies for improving enzyme testing and EFE efficacy in ruminant diets, and to identify proteomic differences between effective and ineffective EFE. A meta-analysis of 20 dairy cow studies with 30 experiments revealed that only a few increased lactational performance and the response was inconsistent. This variability is attributable to several enzyme, feed, animal, and management factors that were discussed in this paper. The variability reflects our limited understanding of the synergistic and sequential interactions between exogenous glycosyl hydrolases, autochthonous ruminal microbes, and endogenous fibrolytic enzymes that are necessary to optimize ruminal fiber digestion. An added complication is that many of the standard methods of assaying EFE activities may over- or underestimate their potential effects because they are based on pure substrate saccharification and do not simulate ruminal conditions. Our recent evaluation of 18 commercial EFE showed that 78 and 83% of them exhibited optimal endoglucanase and xylanase activities, respectively, at 50 °C, and 77 and 61% had optimal activities at pH 4 to 5, respectively, indicating that most would likely act suboptimally in the rumen. Of the many fibrolytic activities that act synergistically to degrade forage fiber, the few usually assayed, typically endoglucanase and xylanase, cannot hydrolyze the recalcitrant phenolic acid-lignin linkages that are the main constraints to ruminal fiber degradation. These factors highlight the futility of random addition of EFE to diets. This paper discusses reasons for the variable animal responses to dietary addition of fibrolytic enzymes, advances explanations for the inconsistency, suggests a strategy to improve enzyme efficacy in ruminant diets, and describes differences among the proteomes of effective and ineffective EFE.

Fibrolytic enzyme and ammonia application effects on the nutritive value, intake, and digestion kinetics of bermudagrass hay in beef cattle.

The objectives were to compare the effect of exogenous fibrolytic enzyme (Biocellulase A20) or anhydrous ammonia (4% DM) treatment on the nutritive value, voluntary intake, and digestion kinetics of bermudagrass (Cynodon dactylon cultivar Coastal) hay harvested after 2 maturities (5- and 13-wk regrowths). Six individually housed, ruminally cannulated Brangus steers (BW 325 ± 10 kg) were used in an experiment with a 6 × 6 Latin square design with a 3 (additives) × 2 (maturities) factorial arrangement of treatments. Each period consisted of 14 d of adaptation and 7, 4, 1, 1, and 4 d for measuring in vivo digestibility, in situ degradability, no measurements, rumen liquid fermentation and passage indices, and rate of solid passage, respectively. Steers were fed hay for ad libitum intake and supplemented with sugarcane molasses and distillers grain (supplement total of 2.88 kg DM/d). Enzyme did not affect the nutritional composition of hay but ammonia treatment decreased hay NDF, hemicellulose, and ADL concentrations and increased the CP concentration particularly for the mature lignified 13-wk hay. The enzyme increased NDF and hemicellulose digestibility of the 5-wk hay but decreased those of the 13-wk hay. Ammoniation decreased intake of hay but increased digestibility of DM, OM, NDF, hemicellulose, ADF, and cellulose and increased the ruminal in situ soluble and potentially digestible fractions and the rate of DM degradation of the 13-wk hay. Also, ammoniation increased the concentrations of ruminal NH3, total VFA, acetate, and butyrate but enzyme treatment did not. Neither enzyme addition nor ammoniation affected rate of liquid and solid passage. In conclusion, ammoniation decreased the concentration of most fiber fractions, decreased the intake of hays, and increased their CP concentration, in vivo digestibility, and in situ degradability at both maturities whereas enzyme application increased fiber digestibility of the 5-wk hay but decreased it in the case of the 13-wk hay.

First report of anthelmintic resistance in gastrointestinal nematodes of sheep from costa rica.

As the prevalence and severity of anthelmintic resistance continue to rise, nematode infections in sheep correspondingly reduce the profitability of the sheep industry. In Costa Rica, sheep production systems are increasing in both number and importance. A field trial study was carried out to detect the level of anthelmintic resistance to albendazole and ivermectin in gastrointestinal nematodes (GIN) of sheep from seven farms in Costa Rica. Resistance was determined using the fecal egg count reduction test (FECRT). Three treatment groups were assessed on each farm: control, albendazole, and ivermectin. Haemonchus spp. (71%), Strongyloides sp. (57%), and Trichostrongylus spp. (43%) presented resistance levels to albendazole, whereas Strongyloides sp. (43%), Haemonchus spp. (29%), and Trichostrongylus spp. (29%) were resistant to ivermectin. Haemonchus spp., Strongyloides sp., and Trichostrongylus spp. were the most resistant GIN to both products. This study suggests that frequency of treatment, exclusive chemical control, and visual estimation of animal weight to calculate dosage may contribute to the high levels of anthelmintic resistance that were observed on the farms analyzed herein.